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使用电纺支架与嵌入细胞外基质水凝胶中的人半月板细胞的新型组合进行半月板组织工程。

Meniscus tissue engineering using a novel combination of electrospun scaffolds and human meniscus cells embedded within an extracellular matrix hydrogel.

作者信息

Baek Jihye, Chen Xian, Sovani Sujata, Jin Sungho, Grogan Shawn P, D'Lima Darryl D

机构信息

Shiley Center for Orthopaedic Research and Education at Scripps Clinic, La Jolla, California; Materials Science and Engineering, University of California, San Diego, La Jolla, California.

出版信息

J Orthop Res. 2015 Apr;33(4):572-83. doi: 10.1002/jor.22802. Epub 2015 Feb 8.

Abstract

Meniscus injury and degeneration have been linked to the development of secondary osteoarthritis (OA). Therapies that successfully repair or replace the meniscus are, therefore, likely to prevent or delay OA progression. We investigated the novel approach of building layers of aligned polylactic acid (PLA) electrospun (ES) scaffolds with human meniscus cells embedded in extracellular matrix (ECM) hydrogel to lead to formation of neotissues that resemble meniscus-like tissue. PLA ES scaffolds with randomly oriented or aligned fibers were seeded with human meniscus cells derived from vascular or avascular regions. Cell viability, cell morphology, and gene expression profiles were monitored via confocal microscopy, scanning electron microscopy (SEM), and real-time polymerase chain reaction (PCR), respectively. Seeded scaffolds were used to produce multilayered constructs and were examined via histology and immunohistochemistry. Morphology and mechanical properties of PLA scaffolds (with and without cells) were influenced by fiber direction of the scaffolds. Both PLA scaffolds supported meniscus tissue formation with increased COL1A1, SOX9, and COMP, yet no difference in gene expression was found between random and aligned PLA scaffolds. Overall, ES materials, which possess mechanical strength of meniscus and can support neotissue formation, show potential for use in cell-based meniscus regeneration strategies.

摘要

半月板损伤和退变与继发性骨关节炎(OA)的发展有关。因此,成功修复或替换半月板的疗法可能会预防或延缓OA的进展。我们研究了一种新方法,即构建嵌入细胞外基质(ECM)水凝胶中的人半月板细胞的对齐聚乳酸(PLA)电纺(ES)支架层,以促使形成类似半月板样组织的新组织。将具有随机取向或对齐纤维的PLA ES支架接种来自血管或无血管区域的人半月板细胞。分别通过共聚焦显微镜、扫描电子显微镜(SEM)和实时聚合酶链反应(PCR)监测细胞活力、细胞形态和基因表达谱。接种后的支架用于制备多层构建体,并通过组织学和免疫组织化学进行检查。PLA支架(有细胞和无细胞)的形态和力学性能受支架纤维方向的影响。两种PLA支架都支持半月板组织形成,COL1A1、SOX9和COMP表达增加,但随机和对齐的PLA支架之间未发现基因表达差异。总体而言,具有半月板机械强度并能支持新组织形成的ES材料显示出在基于细胞的半月板再生策略中应用的潜力。

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